U.S. patent number 4,292,307 [Application Number 06/122,172] was granted by the patent office on 1981-09-29 for vaccine and method for prophylaxis and treatment of clostridioses of animals and poultry.
Invention is credited to Valentina P. Zemlyakova.
United States Patent |
4,292,307 |
Zemlyakova |
September 29, 1981 |
Vaccine and method for prophylaxis and treatment of clostridioses
of animals and poultry
Abstract
A vaccine for prophylaxis and treatment of clostridioses of
animals and poultry is provided which comprises toxoids of
Clostridium perfringens types A, B and D, Cl. oedematiens, and Cl.
septicum, separated from the microbeal mass, a sorbent, and a
solvent, the antigenic activity of said toxoids, expressed in
combining units per ml of the vaccine, being as follows:
Prophylaxis and treatment of clostridioses of animals and poultry
is carried out by giving said animals and poultry parenterally 0.5
to 5 ml of the vaccine, from one to three times, the vaccine
comprising toxoids of Clostridium perfringens types A, B, and D,
Cl.oedematiens, and Cl.septicum, a sorbent and a solvent, the
antigenic activity of said toxoids, expressed in combining units
per ml of the vaccine, being as follows:
______________________________________ toxoid of Cl. perfringens
type A 2-100 toxoid of Cl. perfringens type B 2-100 toxoid of Cl.
perfringens type D 1-80 toxoid of Cl. oedematiens 1-80 toxoid of
Cl. septicum 2-100 sorbent 1-10 mg solvent balance.
______________________________________ The vaccine is universal and
can be used at large industrialized animal and poultry farms to
protect animals and birds from 5-11 causative agents that are
widely spread in nature and are the cause of considerable economic
damage.
Inventors: |
Zemlyakova; Valentina P.
(Minsk, SU) |
Family
ID: |
20724639 |
Appl.
No.: |
06/122,172 |
Filed: |
February 19, 1980 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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942145 |
Sep 14, 1978 |
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Foreign Application Priority Data
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Sep 30, 1977 [SU] |
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2523702 |
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Current U.S.
Class: |
424/239.1 |
Current CPC
Class: |
A61K
39/08 (20130101) |
Current International
Class: |
A61K
9/00 (20060101); A61K 39/08 (20060101); A61K
009/12 (); A61K 039/08 (); A61K 039/116 () |
Field of
Search: |
;424/92 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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288934 |
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Dec 1965 |
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AU |
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895073 |
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May 1962 |
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GB |
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898783 |
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Jun 1962 |
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GB |
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901433 |
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Jul 1962 |
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GB |
|
947912 |
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Jan 1964 |
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GB |
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958574 |
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May 1964 |
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GB |
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Primary Examiner: Rose; Shep K.
Attorney, Agent or Firm: Burns; Robert E. Lobato; Emmanuel
J. Adams; Bruce L.
Parent Case Text
This is a continuation of application Ser. No. 942,145, filed Sept.
14, 1978, now abandoned.
Claims
What is claimed is:
1. A universal vaccine for prophylaxis and treatment of
clostridiosis of animals and poultry, comprising
formaldehyde-detoxicated toxoids of Clostridium perfringens types
A, B, and D, Cl.oedematiens and Cl.septicum separated from the
microbial mass, a sorbent comprised of a aluminum gel and a solvent
comprised of phosphate buffer and physiological salt solution in a
weight ratio of about 1:1, said vaccine being substantially free of
bacterial cells and nonspecific proteins, the antigenic activity of
said components, expressed in combining units per ml of the vaccine
being as follows:
2. A vaccine according to claim 1, which also contains a toxoid of
Cl.tetani, separated from the microbial mass, its antigenic
activity, expressed in combining units per ml of the vaccine, being
1-80.
3. A vaccine according to claim 2, which also contains toxoids of
Cl.botulinum of at least two of the types A, B, C, D, E, and F,
separated from the microbial mass, their antigenic activity,
expressed in combining units per ml of the vaccine, being as
follows:
4. A vaccine according to claim 1, containing a mixture of a
phosphate buffer and a physiological salt solution, taken in the
weight ratio of 1:1, as the solvent.
5. A vaccine according to claim 1, containing alumina gel as the
sorbent.
6. A method for prophylaxis and treatment of clostridiosis of
animals, and poultry, comprising treating said animals and poultry
with a vaccine in a dose of 0.5 to 5 ml, from one to three times,
the vaccine comprising toxoids of Clostridium perfringens types A,
B, and D, Cl.oedematiens, and Cl. septicum, separated from the
microbial mass, a sorbent comprised of aluminum gel and a solvent
comprised of phosphate buffer and physiological salt solution in a
weight ratio of about 1:1, said vaccine being substantially free of
bacterial cells and nonspecific proteins the antigenic activity of
said toxoids, expressed in combining units per ml of the vaccine,
being as follows:
with subsequent re-vaccination.
7. A method for prophylaxis and treatment of clostridiosis
according to claim 6, in which said animals and poultry are given
said vaccine also containing a toxoid of Cl.tetani, separated from
the microbial mass, its antigenic activity, expressed in combining
units per ml of the vaccine, being 1-80.
8. A method for prophylaxis and treatment of clostridiosis
according to claim 6, in which said animals and poultry are given
the vaccine also containing toxoids of Cl.botulinum of at least two
of the types A, B, C, D, E, and F, separated from the microbial
mass, their antigenic activity, expressed in combining units per ml
of the vaccine, being as follows:
9. A method for prophylaxis and treatment of clostridiosis
according to claim 6, in which said animals and poultry are given
said vaccine parenterally.
10. A method for prophylaxis and treatment of clostridiosis
according to claim 6, in which said animals and poultry are given
the vaccine in an aerosol form.
11. A vaccine according to claim 1 which has an expiration time of
about 5 years.
12. A method according to claim 1 wherein immunity to said
clostridiosis lasting about 1 to 2 years is provided.
Description
FIELD OF THE INVENTION
The present invention relates to veterinary microbiology and more
particularly it relates to a vaccine for prophylaxis and treatment
of clostridiosis in animals and birds.
BACKGROUND OF THE INVENTION
A a polyvalent antigenic vaccine against clostridioses of animals,
is known comprising antigens that are anacultures of Cl.
perfringens types B, C, and D, Cl. oedematiens, Cl. septicum, Cl.
tetani, and Cl. chauvoei, a mineral oil, a nonionic lipophilic
emulsifying agent, and a nonionic hydrophilic emulsifying
agent.
The presence of microbial cells in the vaccine produces a negative
effect on detoxification and purification of the preparation to
reduce its antigenic and immunogenic power, to increase the
reactogenecity, and produces mainly antimicrobial immunity in the
vaccinated animals.
A precipitated toxoid-vaccine is also known which contains a toxoid
of Cl. perfringens type A, anacultures Cl. perfringens types B, C,
and D, Cl. oedematiens, and Cl. septicum separated from the
microbial mass. Alumina gel is used as an adsorbent. The activity
of the toxoid of Cl. perfringens type A is measured in combining
units per ml of the vaccine and is equal to 20 units.
The number of combining units characterizes the antigenic and
immunogenic properties of the toxoid.
A combining unit is the quantity of the test toxoid (at various
stages of its preparation), capable of combining with one antitoxic
unit of specific serum. The specific activity of toxoids, expressed
in combining units, is determined by the method proposed by G. P.
Cherkas (1958) and I. M. Khaustova (1959).
The activity of each toxin of the starting cultures that are
constituents of said vaccine is determined by the MLD (minimum
lethal dose) and is, on an average, 400-1000 MLD in one ml of the
vaccine (for albino mice).
The antigenic and immunogenic properties of the preparation are
insufficiently strong, and it exhibits, mainly, antimicrobial
immunity and a very weak antitoxic immunity against Clostridium
perfringens types B, C, and D, Cl. oedematiens and Cl.
septicum.
The vaccinated animals develop immunity very slowly, and it lasts
for a short time: the immunity sharply weakens in 1-2 months, which
involves repeated vaccinations with increased doses of the
preparation.
A another vaccine, namely ditoxoid, is also known which is intended
for specific prophylaxis of enterotoxemia of calves. The
preparation includes toxoids of Clostridium perfringens types A and
D, separated from the microbial mass, their antigenic activity
being 100 combining units of each toxoid per ml, as well as a
sorbent and a solvent. The sorbent used in the vaccine in alumina
gel and the solvent, a physiological salt solution.
Said preparation cannot be used for treatment and prophylaxis of
diseases caused by a combination of widely spread natural
microorganisms of the genus Clostridium since it contains toxoids
effective only against two causative agents.
SUMMARY OF THE INVENTION
The principal object of the invention is to improve the antigenic
and immunogenic properties of the vaccine and to provide strong
antitoxic immunity in vaccinated animals.
Another object of the invention is to provide a universal and
stable vaccine against a combination of most widely spread
microorganisms of the genus Clostridium.
The present invention resides in that a vaccine for prophylaxis and
treatment of clostridiosis of animals and poultry comprises toxoids
of Clostridium perfringens types A, B, and D, Cl. oedematiens, Cl.
septicum, separated from the microbial mass, a sorbent and a
solvent, the antigenic activity of said toxoids, expressed in
binding units per ml of the vaccine, being as follows:
______________________________________ toxoid of Cl. perfringens
type A 2-100 toxoid of Cl. perfringens type B 2-100 toxoid of Cl.
perfringens type D 1-80 toxoid of Cl. oedematiens 1-80 toxoid of
Cl. septicum 2-100 sorbent 1-10 mg solvent balance.
______________________________________
Said vaccine is used against the following diseases of cattle,
horses, sheep, pigs, commercial animals and poultry: gas gangrene,
malignant edema, endometritis, vaginitis, mastitis, miscarriage,
bradzot of sheep, and enterotoxemia of the younger livestock, that
are produced by said caustive agents.
It is not recommended to use the vaccine in which the antigenic
activity of its components is below the minimum doses since the
vaccine will fail to produce the antitoxic immunity in the
vaccinated animals and birds at the protective level.
Nor is it recommended to use the vaccine in which the antigenic
activity of its components is higher than their maximum doses,
since the production of antitoxins in the vaccinated animals is
inhibited in excess (antitoxic immunity).
It is recommended that the vaccine also contain a toxoid of Cl.
tetani, separated from the microbial mass, its antigenic activity,
expressed in combining unit per ml of the vaccine, being 1-80.
The proposed vaccine is used against infectious enterotoxemia in
young animals, mainly newborns of all agricultural animals and
poultry, in female adults of cattle, horses, sheep, pigs, against
gynaecological diseases (endometritis, vaginitis, mastitis), wound
complications, gas gangrene, malignant edema, bradzot, of sheep,
that are caused by said combination of microorganisms of the genus
Clostridium. It is recommended that the vaccine also contain
toxoids of Cl. botulinum of at least two of the types A, B, C, D,
E, and F, separated from the bacterial mass, their antigenic
activity, expressed in combining units per ml of the vaccine, being
as follows:
______________________________________ toxoid of Cl. botulinum type
A 2-100 toxoid of Cl. botulinum type B 0.2-80 toxoid of Cl.
botulinum type C 0.2-80 toxoid of Cl. botulinum type D 0.2-80
toxoid of Cl. botulinum type E 0.2-80 toxoid of Cl. botulinum type
F 0.2-80 ______________________________________
The proposed vaccine is used for prophylaxis and treatment of
forage toxicoinfection, anaerobic enterotoxemia, gynaecological
diseases and wound complications.
The incorporation into the vaccine of toxoids of Cl. tetani and Cl.
botulinum types A, B, C, D, E, and F is explained by the necessity
of prophylaxis and treatment of clostridiosis caused by said
microorganisms.
The selection of the minimum and maximum doses of antigenic
activity of toxoids of Cl. botulinum and Cl. tetani in the proposed
vaccine is governed by the same reasons as specified above for
toxoids of Cl. perfringens types A, B, and D, Cl. oedematiens, and
Cl. septicum.
It is recommended that the vaccine contain alumina gel as the
sorbent. This sorbent ensures complete sorption of the toxoids and
does not produce any harmful effect on animals.
It is recommended to use a mixture of a phosphate buffer and a
physiological salt solution, taken in the weight ratio of 1:1, as
the solvent.
The invention also resides in a method for prophylaxis and
treatment of clostridiosis of animals and poultry, consisting in
that said animals and poultry are given the vaccine in doses from
0.5 to 5 ml, from one to three times, the vaccine comprising
toxoids separated from the microbial cells of Clostridium
perfringens, types A, B, and D, Cl. oedematiens, Cl. septicum, a
sorbent and a solvent, the antigenic activity of said toxoids,
expressed in combining units per ml of the vaccine, being as
follows:
______________________________________ toxoid of Cl. perfringens,
type A 2-100 toxoid of Cl. perfringens type B 2-100 toxoid of Cl.
perfringens type D 1-80 toxoid of Cl. oedematiens 1-80 toxoid of
Cl. septicum 2-100 sorbent 1-10 mg solvent balance
______________________________________
with subsequent re-vaccination.
To maintain the immunity at the protective level, the animals
should be re-vaccinated once a year or for special indications
(infection outbreak).
It is recommended that the animals and poultry be given the vaccine
also containing a toxoid of Cl. tetani, separated from the
microbial cells, its antigenic activity, expressed in combining
unit per ml of the vaccine, being 1-80.
It is also recommended that the animals and poultry be given the
vaccine containing a toxoid of Cl. botulinus (separated from the
microbial cells) of at least two types out of the types A, B, C, E,
and F, their antigenic activity, expressed in combining units per
ml of the vaccine, being as follows:
______________________________________ toxoid of Cl. botulinus type
A 2-100 toxoid of Cl. botulinus type B 0.2-80 toxoid of Cl.
botulinus type C 0.2-80 toxoid of Cl. botulinus type F 0.2-80
toxoid of Cl. botulinus type F 0.2-80
______________________________________
It is recommended to given the vaccine parenterally or in an
aerosol form.
DETAILED DESCRIPTION OF THE INVENTION
The proposed vaccine is a liquid preparation that separates on
standing into a colorless supernatant liquid and a white,
pale-brown, or yellowish precipitate. When shaken, the vaccine
gives a homogeneous suspension free from flakes or extraneous
impurities.
The vaccine has been tested on smaller laboratory animals and on
agricultural animals such as cattle, sheep, pigs, camels, horses,
etc., on newborn calves, gilts, lambs, poultry (e.g. hens), and
commercial animals (minks).
Observation of the animals has shown that the active formation of
intense immunity stimulated the production of a high titer of
toxoids to each component of the vaccine, and the vaccinated
animals developed immunity (in primary immunization) in 10-25 days
and (after re-vaccination) in 5-10 days. The length and activity of
the immunity produced by the proposed vaccine were studied on 100
animals that were infected with the corresponding virulent culture
or a toxin, which is a constituent of the proposed vaccine,
1,2,4,6,10,12 months after the vaccination and 2,4, and 5 years
after the vaccination. Non-immunized animals, and also animals who
had the corresponding disease in the past, were used as controls.
The animals were selected into the control group according to the
analogue principle.
The animals were immunized in accordance with the immunization
methods developed by the inventors for each species of animals and
poultry.
In cases where one or several said causative agents were revealed,
all profitable healthy cattle, calves, pigs, sheep, horses, camels,
poultry, minks, and other animals were given a prophylactic
vaccination irrespective of the pregnancy term. At affected farms,
where the preparation was used for the first time, the cattle,
horses, camels, pigs and sheep were vaccinated from one to three
times. The vaccine was given in doses from 0.5 to 5 ml,
parenterally or in an aerosol form. The animals and poultry were
re-vaccinated 3-6 months following the first vaccination, at the
time of intense reproduction, or for special indications (outbreaks
of forage poisoning, etc.).
In cases of urgent prophylaxis of anaerobic enterotoxemia of
newborns, when the vaccination of the mother is useless, the
newborns of non-vaccinated mothers are given the vaccine. The
vaccination is done in two injections.
The vaccine does not produce any harmful effect on the vaccinated
animal or the posterity. It is harmfull and areactogenic. The
newborn animals are healthy, strong, viable, and gain weight
quickly. The vaccine proves quite effective against the diseases
caused by Clostridium perfringens types A, B, and D, Cl.
oedematiens, Cl. septicum, Cl. titani, and Cl. botulinum. The
efficacy is hundred percent. At the same time, all animals in the
control group (100 percent) were affected by the disease and from
80 to 100 percent animals perished.
The vaccine remains effective up to five years.
The proposed vaccine for prophylaxis and treatment of clostridiosis
in animals and poultry is prepared as follows.
A growth medium containing sources of nitrogen, carbon, phosphorus
and mineral salts is inoculated with separate cultures of
Clostridium perfringens types A, B, and D, Cl. oedematiens, Cl.
septicum, Cl. tetani, and Cl. botulinum types A, B, C, E, and F.
The cultures are grown under anaerobic conditions at a temperature
of 35.degree.-38.degree. C., the pH of the medium being 5-10. The
time of cultivation depends on the species and type of the
microorganisms and averages from two hours to ten days. At the end
of the cultivation period, a culture suspension of each species and
type is separated into the culture filtrate and the microbial
mass.
The activity of toxins, expressed in MLD per ml for albino mice is
as follows:
______________________________________ toxin of Cl. perfringens
type A 800-1200 toxin of Cl. perfringens type B 1000-3000 toxin of
Cl. perfringens type D 1000-3000 toxin of Cl. oedematiens
12,000-20,000 toxin of Cl. septicum 800-1000
______________________________________
The culture filtrate containing the toxin is separated and passed
through a Seitz filter. Then, the toxin is partially detoxicated by
adding 0.4-0.5 percent by volume of a 40 percent formaldehyde
solution into the culture filtrate at a temperature of
37.degree.-38.degree. C., the pH of the medium being 7-7.2.
Formaldehyde is added in portions, once or twice within 1-20 days,
depending on the species and the type of the microorganism
producing the toxin. The only exception are toxins of Cl.
perfringens type B, Cl. oedematiens and Cl. septicum, that are
fully attenuated with formaldehyde within 1 to 30 days to give the
toxoid.
The partially attenuated toxin or toxoid is then concentrated and
purified by precipitating with a 45-60 percent solution of ammonium
sulphate or sodium hexametaphosphate in the presence of IN
hydrochloric acid. The precipitated toxin or toxoid is dissolved in
a mixture of a phosphate buffer and physiological salt solution
taken in the weight ratio of 1:1. The obtained solution is
sterilized by passing through Seitz filters. Adding a 40 percent
formaldehyde solution to the partly attenuated toxin at a
temperature of 37.degree.-38.degree. C. converts it into the
toxoid.
The obtained toxoid is tested for specific harmlessness on guinea
pigs, and its antigenic activity, expressed in combining units, is
determined. The toxoid is also tested for sterility.
Thus obtained monotoxoids, separated from the microbial cells, are
mixed together and adsorbed on aluminium hydroxide gel.
If necessary, a preservative may be added to the prepared vaccine,
such as a solution of sodium merthiolate (1:10,000).
The prepared vaccine is tested for sterility, toxicity, Al.sub.2
O.sub.3 content, and immunogenecity.
The proposed vaccine possesses high antigenic and immunogenic
properties and produces in vaccinated animals an effective
antitoxic immunity to all said causative agents. The immunity
persists from 1 to 2 years. The immunity to specific agents can
last to 5 years. The vaccine dose is reduced two times. The titers
of antitoxins in animals toward all monotoxoids increase 8-10
times, and the antitoxins are produced twice as fast. Following the
primary immunization, the animals are re-vaccinated only once in
their lifetime or only for special epizootic indications (an
intervals from 2 to 5 years). The intensity of the immunity is
15-20 times higher than that attained with the known polyvalent
toxoid-vaccine. The preparation is harmless and areactogenic for
both vaccinated animals and man (milk and meat of vaccinated
animals). The time of expiration of the vaccine has increased to
five years.
Compared with the known preparations, the proposed vaccine is free
from ballasts (microbial cells, nonspecific proteins). The activity
of the vaccine is determined by the combining units and is
controlled at all stages of its production and use, which
guarantees efficacy of the vaccine. The method of testing the
vaccine, especially for immunogenecity and safety has been improved
as well.
The use of the proposed vaccine for prophylactic purposes at
affected farms, especially at large industrialized complexes, has
proved its universal properties since it can effectively be used
for vaccination of all animals or poultry against 5-11 widely
spread natural causative agents, the vaccination dose being small.
The vaccine protects females from forage poisoning, wound
complications, and complications following the delivery. The
vaccine can also be used for prophylaxis and treatment of saplings
from anaerobic enterotoxemia, through the agency of colostrum of
their mothers, guarantees complete cure of diseased animals and
specific prophylxis of the mothers themselves against said
causative agents, the efficacy being 100 percent. Rare cases of
infections are not serious and the symptomatic treatment ensures
rapid cure. Animals borne from vaccinated mothers are viable,
strong, and rapidly gain weight. The use of the proposed vaccine at
affected animal farms ensures rapid eradication of the diseases. At
large animal breeding farms, the vaccine ensures the formation of
the herd free from 5-11 widely spread natural causative agents that
otherwise are the cause of considerable loss.
For a better understanding of the invention, the following examples
of its practical embodiment are given by way of illustration.
EXAMPLE 1
Ten liters of a casein growth medium sterilized at a temperature of
112.degree. C. for thirty minutes are inoculated with the culture
Clostridium perfringens type A, taken in a quantity of 2 percent of
the growth medium volume. The cultivation is carried out under
anaerobic conditions at a temperature of 38.degree. C. for 2-10
hours at pH of the medium of 5-10. The same casein medium is used
for parallel cultivation of Cl. perfringens types B, D, Cl.
oedematiens, Cl. septicum, Cl. tetani, and Cl. botulinum types A,
B, C, D, E, and F. All cultures are grown separately by the
described method except that the time of cultivation for each
culture is from 2 hours to 10 days.
At the end of cultivation, 12 liters of the suspension of each
culture are separated into the culture filtrate and the microbial
mass. The culture filtrate (11.5 liters) containing the toxin is
separated and passed through a Seitz filter. The toxin is then
detoxicated with a 40 percent formaldehyde solution.
The toxin of Clostridium perfringens type A is partly detoxicated
with a 40 percent formaldehyde solution at a temperature of
37.degree.-38.degree. C. for 14 days. Formaldehyde is added in two
portions (0.2 percent by volume in each portion) at a 24-hour
interval.
The toxin of Cl. perfringens type B is detoxicated and converted
into the corresponding toxoid with a 40 percent solution of
formaldehyde at pH 7.0-7.2, at a temperature of
37.degree.-38.degree. C., for 1-3 days. The entire portion of
formaldehyde (0.4-0.5 percent by volume) is added at a time.
The toxin of Cl. perfringens type D is detoxicated partly with a 40
percent solution of formaldehyde at pH of the medium of 7-7.2, at a
temperature of 37.degree.-38.degree. C., for 1-30 days, the
required quantity of formaldehyde being added in two portions (0.2
percent by volume in each portion) at a 1-2 day interval.
The toxins of Cl. oedematiens and Cl. septicum are detoxicated and
converted into the corresponding toxoids by the same procedure as
described for Cl. perfringens type B.
The attenuated toxin or toxoid obtained from the culture filtrate
(11.6 liters) is purified and concentrated by precipitating with a
25 percent solution of sodium hexametaphosphate, taken in a
quantity of 0.5-1 percent by volume, in the presence of IN
hydrochloric acid at pH of 3.0-3.8.
The precipitated toxin or toxoid (100 g) is dissolved in three
liters of a mixture of a phosphate buffer and a physiological salt
solution (1:1), and the obtained solution is sterilized by passing
it through Seitz filters.
The resultant products are concentrated solutions (separated from
microbial cells) of toxoids of Cl. perfringens type B, Cl.
oedematiens, Cl. septicum, and concentrated solutions of partly
detoxicated toxins of Cl. perfringens types A and D.
The concentrated solution of partly detoxicated Cl. perfringens
type A is converted into the corresponding toxoid by adding 0.2
percent by volume of a 40 percent formaldehyde solution in the
course of 14-20 days.
The partly detoxicated toxin of Cl. perfringens type D is converted
into the toxoid by a procedure similar to that described for Cl.
perfringens type A.
The obtained monotoxoids, separated from microbial cells, are
tested for sterility, toxicity, specific properties, and antigenic
activity.
The antigenic activity of concentrated solution of each toxoid,
separated from microbial cells, expressed in combining units per ml
of solution, is as follows:
______________________________________ toxoid of Cl. perfringens
type A 350 toxoid of Cl. perfringens type B 300 toxoid of Cl.
perfringens type D 500 toxoid of Cl. oedematiens 500 toxoid of Cl.
septicum 200 ______________________________________
The concentrated solutions separated from microbial cells, prepared
as described above, are diluted with a physiological salt solution
(0.85 percent sodium chloride solution) to obtain solutions having
the following antigenic activities expressed in the combining units
per ml of the vaccine:
______________________________________ toxoid of Cl. perfringens
type A 2 toxoid of Cl. perfringens type B 2 toxoid of Cl.
perfringens type D 1 toxoid of Cl. oedematiens 1 toxoid of Cl.
septicum 2 ______________________________________
To prepare 2.5 l of the vaccine according to the invention, 200 ml
of dilute solutions of each monotoxoid, having said antigenic
activity, are mixed together, and alumina gel is added (400 ml as
Al.sub.2 O.sub.3).
The finished vaccine contains the following components having the
following antigenic activity, expressed in combining units per ml
of the vaccine:
______________________________________ toxoid of Cl. perfringens
type A 2 toxoid of Cl. perfringens type B 2 toxoid of Cl.
perfringens type D 1 toxoid of Cl. oedematiens 1 toxoid of Cl.
septicum 2 alumina gel (as Al.sub.2 O.sub.3) 1 ml mixture of
phosphate buffer and physiological salt solution (1:1) balance.
______________________________________
The finished vaccine is tested by known methods for completeness of
sorption, sterility, toxicity, and immunogenecity (on mice). The
vaccine is sterile, harmless, and immunogenic.
In order to prevent development of enterotoxemia in young pigs and
calves at farms where 100 percent of the animals were affected with
the disease (against the background of dyspepsia), the mortality
rate being 50-80 percent, all adult female cattle and pigs were
immunized with the vaccine.
The development of the disease in the young stock was as follows
profuse diarrhea on the 2nd or 3rd day following the birth,
sometimes with blood in the feces, the animals abstained from food,
lost weight quickly and died within 2-3 days. In female animals,
endometrites and mastites were observed.
The vaccine was used to immunize 100 pregnant cows and 150 pigs.
The vaccine was given parenterally, 5 ml in two injections. The
control group contained 20 cows and 50 pigs. The immunized animals
delivered 95 calves and 1200 gilts. Fifty young calves got the
disease in a mild form and ten of them perished. The affected gilts
were 50 as well, and 25 of them perished.
All animals in the control group were affected by the disease, and
a hundred percent mortality was observed with the calves and gilts
born from the control animals.
EXAMPLE 2
The vaccine contains the following components, their antigenic
activity, expressed in combining units per ml, being as
follows:
______________________________________ toxoid of Cl. perfringens
type A 10 toxoid of Cl. perfringens type B 10 toxoid of Cl.
perfringens type D 5 toxoid of Cl. oedematiens 4 toxoid of Cl.
septicum 10 alumina gel 5 mg (as Al.sub.2 O.sub.3) mixture of
phosphate buffer and physiological salt solution (1:1) balance.
______________________________________
The vaccine is prepared by a procedure similar to that described in
Example 1.
The finished preparation is sterile, harmless, and immunogenic. It
is used for prophylaxis and treatment of calves and gilts with
anaerobic enterotoxemia against the background of dyspepsia. The
clinic of the disease is similar to that described in Example
1.
The vaccine was used to immunize 100 cows and 230 pigs in
pregnancy. The vaccine was given pareneterally in a dose of 5 ml in
two injections. The control group contained 20 cows and 30 pigs.
The immunized animals delivered 97 calves and 1796 gilts. The
disease did not develop, and none of the young stock perished. The
non-immunized cows gave 18 calves, all of them had the grave form
of the disease and 12 calves perished. The non-immunized pigs gave
205 gilts, 200 of them got the disease and 150 perished.
EXAMPLE 3
The vaccine contains the following components, their antigenic
activity, expressed in combining unit per ml, being as follows:
______________________________________ toxoid of Cl. perfringens
type A 100 toxoid of Cl. perfringens type B 100 toxoid of Cl.
perfringens type D 80 toxoid of Cl. oedematiens 80 toxoid of Cl.
septicum 100 alumina gel 10 mg (as Al.sub.2 O.sub.3) mixture of
phosphate buffer and physiological salt solution (1:1) balance.
______________________________________
The proposed vaccine is prepared by the method described in Example
1.
The preparation is sterile, harmless, immunogenic and can be used
for prophylaxis and treatment of young animals (calves, pigs) with
anaerobic enterotoxemia against the background of dyspepsia, the
clinic of the disease being similar to that described in Example
1.
The proposed vaccine was used to immunize 50 cows and 100 pigs in
pregnancy. The vaccine was given in two injections in a dose of 5
ml. The control group contained 20 cows and 50 pigs. The immunized
animals produced 50 calves and 960 gilts. 25 gilts developed the
disease and 10 of them perished. 25 calves developed the disease in
the mild form and ten of them perished. 200 young pigs were
slightly affected and 50 of them perished.
One hundred percent of the animals in the control group got the
disease.
All animals delivered by the non-immunized cows and pigs in the
control group developed the disease and perished.
EXAMPLE 4
The vaccine contains the following components their antigenic
activity, expressed in combining units per ml, being as
follows:
______________________________________ toxoid of Cl. perfringens
type A 2 toxoid of Cl. perfringens type B 2 toxoid of Cl.
perfringens type D 1 toxoid of Cl. oedematiens 1 toxoid of Cl.
septicum 2 toxoid of Cl. tetani 1 alumina gel 1 mg (as Al.sub.2
O.sub.3) mixture of phosphate buffer and physiological salt
solution (1:1) balance. ______________________________________
The finished preparation is tested by the known methods for
sterility, toxicity, and immunogenecity.
The vaccine is sterile, harmless, and immunogenic.
In order to prevent and treat anaerobic enterotoxemia of young
cattle, pigs, sheep, horses, of endometritis, vaginitis, mastitis,
wound complications, malignant edema, bradzot, of sheep, etc., the
animals at farms affected with the diseases were immunized with the
proposed vaccine. The disease among females was characterized by
purulent discharges from the uterus and vagina, mastitis,
miscarriages, malignant edema, about 50 to 70 percent animals being
affected by the diseases. The disease in the young animals
manifested in grave anaerobic enterotoxemia, the clinic being the
same as described in Example 1, associated with grave nerve
affections and resulting in 100 percent mortality among the
newborns who perished within the first 3 days.
The vaccine was given to 200 cows, 219 pigs, 120 sheep and 80
horses in pregnancy. The preparation was given in two injections in
a dose of 5 ml which was injected from one to three times,
depending on the epizootic situation. The control group contained
20 pigs, 20 sheep, 20 horses and 20 cows. The immunized 150 pigs
delivered 1350 gilts, the horses gave 60 foals, the sheep gave 150
lambs, and the cows gave 198 calves. The mild form of the disease
was observed in 79 gilts, and the grave form in 70 gilts; 28 of
them perished. The mild form developed in 30 foals and two of them
perished. 60 lambs got diseased and 5 of them perished. The
prophylactic use of the proposed vaccine in female animals to
prevent endometritis, vaginitis, miscarriages and wound
complications was effective in 75 percent. cases. The diseases were
not eradicated in the control group.
Hundred percent young animals born from the non-immunized mothers,
developed the disease and perished.
EXAMPLE 5
The vaccine contains the following components, their antigenic
activity, expressed in combining units per ml., being as
follows:
______________________________________ toxoid of Cl. perfringens
type A 10 toxoid of Cl. perfringens type B 10 toxoid of Cl.
perfringens type D 5 toxoid of Cl. oedematiens 4 toxoid of Cl.
septicum 10 toxoid of Cl. tetani 5 alumina gel 5 mg (as Al.sub.2
O.sub.3) mixture of phosphate buffer and physiological salt
solutions (1:1) balance. ______________________________________
The vaccine is prepared by a procedure similar to that described in
Example 1. The finished preparation is sterile, harmless, and
immunogenic, and it can be used for prophylaxis and treatment of
young livestock (lambs, foals, calves, gilts, etc.) and female
adults. The vaccine is effective against the diseases indicated in
Example 4.
The proposed vaccine was used to immunize 242 pigs, 200 sheep, 150
horses, and 400 cattle in pregnancy. The preparation was given
parenterally in a dose of 5 ml (from 1 to 3 times), depending on
the epizootic situation. The control group contained 24 animals of
each species. The immunized animals gave 1630 gilts, 180 lambs, 80
foals, and 389 calves. All young animals survived and none of them
got the disease. The same picture was observed with the adults
immunized with the vaccine. The diseases were not eradicated among
the animals in the control group. Hundred percent animals born from
the non-immunized mothers got the disease and perished.
EXAMPLE 6
The vaccine contains the following components, their antigenic
activity, expressed in combining units per ml, being as
follows:
______________________________________ toxoid of Cl. perfringens
type A 10 toxoid of Cl. perfringens type B 10 toxoid of Cl.
perfringens type D 5 toxoid of Cl. oedematiens 4 toxoid of Cl.
septicum 10 toxoid of Cl. tetani 5 toxoid of Cl. botulinum type A
10 toxoid of Cl. botulinum type B 2 toxoid of Cl. botulinum type C
2 toxoid of Cl. botulinum type D 2 toxoid of Cl. botulinum type E 2
toxoid of Cl. botulinum type F 2 alumina gel 5 mg (as Al.sub.2
O.sub.3) mixture of phosphate buffer and physiological salt
solution (1:1) balance. ______________________________________
The vaccine is prepared by a procedure similar to that described in
Example 1.
The finished preparation is sterile, harmless, and immunogenic.
The vaccine was used to immunize commercial animals and poultry in
order to prevent and treat forage toxicoinfections, wound
complications, gas gangrene, post-labour complications, and
enterotoxemia at animal and poultry farms affected with the
diseases. The affection of the animals and poultry was
characterized by signs of poisoning and quick lethal outcomes.
Said vaccine was used to immunize 200 minks, 500 birds and 150
pigs. The preparation was given to minks in a single injection of
1-2 ml with subsequent re-vaccination, or in aerosol form (2-3 ml).
Pigs were immunized twice with 5 ml doses. The parenteral dose to
hens and ducks was 0.5-2 ml, which was given 1 or 2 times,
depending on the epizootic situation, or in aerosol form (1-3 ml).
The control group contained 100 minks, 250 birds and 20 pigs. The
immunized minks delivered 150 cubs and the pigs delivered 1350
gilts without any signs of the disease in the mothers and young
animals, and all of them survived. A mild form of the disease was
observed with 10-15 percent immunized birds. The disease was only
transient (1-2 days), without lethal outcomes.
The disease affected 95 percent of the minks, poultry and pigs,
including the mothers and young animals, in the control group, and
95 percent of them perished.
EXAMPLE 7
The vaccine contains the following components their antigenic
activity, expressed in combining units per ml, being as
follows:
______________________________________ toxoid of Cl. perfringens
type A 10 toxoid of Cl. perfringens type B 10 toxoid of Cl.
perfringens type D 5 toxoid of Cl. oedematiens 4 toxoid of Cl.
septicum 10 toxoid of Cl. tetani 5 toxoid of Cl. botulinum type A
10 toxoid of Cl. botulinum type C 2 alumina gel 5 mg (as Al.sub.2
O.sub.3) mixture of phosphate buffer and physiological salt
solution (1:1) balance. ______________________________________
The proposed vaccine is prepared by the method described in Example
1.
The finished preparation is sterile, harmless, and immunogenic. It
is used for prophylaxis and treatment of commercial animals and
poultry and is effective against the diseases indicated in Example
6.
Said vaccine was used to immunize 150 minks and 300 birds. The
minks were given the vaccine in a single injection of 1-2 ml with
subsequent re-vaccination, or in aerosol form (2-3 ml). Poultry
(hens and ducks) were given 0.5-2 ml of the vaccine, from 1 to 2
times, depending on the epizootic situation. The control group
contained 50 minks and 100 birds. The immunized minks delivered 100
cubs without any signs of the disease. All of them survived. The
posterity of the immunized poultry was not affected by the disease
either, and all chicks survived. About 90-100 percent of the
non-immunized minks and poultry developed the diseases and
perished.
* * * * *